1. Volume 18, Issue 10, Pages 1320-1330 (October 2011)
Promysalin, a Salicylate-Containing Pseudomonas putida Antibiotic, Promotes Surface Colonization and Selectively Targets Other Pseudomonas 
Wen Li, Paulina Estrada-de los Santos, Sandra Matthijs, Guan-Lin Xie, Roger Busson, Pierre Cornelis, Jef Rozenski, René De Mot 
Chemistry & Biology 
Volume 18, Issue 10, Pages (October 2011)
DOI: /j.chembiol
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2. Chemistry & Biology 2011 18, 1320-1330DOI: (10. 1016/j. chembiol. 2011
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3. Figure 1
Main Fragments Obtained by Collision-Induced Fragmentation in a Mass Spectrometer
Hydrogens in bold are exchangeable; the underlined hydrogen is partially exchanged in deuterated methanol. Spectra are shown in Figure S1.
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4. Figure 2 Structure of Promysalin
Numbering of carbon atoms for NMR assignments is indicated. In addition to promysalin, three related minor metabolites (dihydropromysalin, deoxypromysalin, dihydrodeoxypromysalin) were identified in cell extracts (see Figure S3).
Chemistry & Biology  , DOI: ( /j.chembiol )
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5. Figure 3
Phenotypes of P. putida RW10S1 and Representative Derived Strains
Antagonistic activity with P. stutzeri as overlay indicator (A) and swarming ability on media with different agar concentrations (0.5%–1.5%) (B) of RW10S1, promysalin biosynthetic mutants M-17 (insertion in ppgH) and M-63 (insertion in ppgO), and regulatory mutant M-50 (insertion in gacS). Minus and plus signs refer to mutants and complemented strains, respectively, using the promysalin biosynthetic region (cosmid pCMPG6110) or the subcloned gacS gene (shuttle vector pCMPG6113). Antibacterial activity of P. putida KT2440 and P. fluorescens OE 28.3 carrying pCMPG6110 is also shown (A). No swarming activity was conferred upon these heterologous producers (not shown). All mutants are described in Table S2.
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6. Figure 4 Organization of the Promysalin Biosynthetic Gene Cluster
Gray shading (inset) refers to predicted ppg gene functions (see also Table 2 and Figure 5). Plasposon insertion sites generating mutants without antagonistic activity (solid line) or mutants with reduced antagonistic activity (dashed line) are indicated. The following genes overlap: ppgAB, ppgCDE, ppgGH, and ppgKLM. A potential transcriptional ρ-independent terminator (black diamond) is indicated. Details of a putative target for translational repressor proteins of the Gac/Rsm system (black triangle) and of the syntenous regions flanking the 15.7 kb biosynthetic cluster (wavy lines) are shown in Figure S4. The agarose electrophoresis panel shows the transcript analysis by RT-PCR with primer couples for each combination of adjacent convergent gene pairs from left lane (ppgA-ppgB) to right lane (ppgN-ppgO). Size markers (bp) are indicated (see Supplemental Experimental Procedures for expected amplicon sizes). FA, fatty acid; SA, salicylic acid; Pro, proline.
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7. Figure 5 Proposed Pathway for Promysalin Biosynthesis
The promysalin core is assembled from proline, myristic acid, and salicylic acid. Chorismic acid serves as a salicylic acid precursor. The proposed scheme is supported by the identification of intermediates (2–6) arising in specific mutants (see Figure S6) and minor compounds detected in the wild-type cells (1–3; see Figure S3). Fatty acid biosynthesis involving PpgC, PpgD, PpgE, and PpgK is not shown. PpgA probably is a carrier for an unspecified amino acid-containing intermediate. The remaining gene products (PpgB, PpgI, PpgL) cannot be assigned to the one of the steps marked with dashed arrows.
Chemistry & Biology  , DOI: ( /j.chembiol )
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